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Description

Unique blend of highly-efficient MyTaq HS DNA Polymerase and a proprietary proofreading enzyme that combine to give increased target affinity for use with challenging templates and inhibitor-rich samples.

Product Highlights

Robust - enzyme blend and buffering system promotes reliable amplification of the most challenging and complex targets, even in the presence of inhibitors

Product Description

MyFi™ has been developed to give reliable amplification of targets up to 10 kb from challenging and complex targets, even in the presence of PCR inhibitors. MyFi is therefore ideal for amplification of cDNA libraries, complex genomic fragments and GC-rich targets. Additionally, MyFi shows improved tolerance to PCR inhibitors, thereby enabling reliable detection from samples from which DNA is difficult to purify. Furthermore, its unique buffering system and enzyme blend promote highly sensitive amplification of even very low-copy number targets. The proofreading ability of MyFi allows all PCR products to be cloned. The inclusion of MyTaq HS means MyFi generates PCR products with 3’-A overhangs, which is perfect for TA cloning. MyFi has the added convenience of room temperature reaction assembly, to avoid non-specific amplification and primer-dimer formation.

Download the PCR Enzyme Guide with detailed product descriptions and performance data to help you choose the best product for your research

We commonly use the Bioline MyFi polymerase, and it works better than any other polymerase. MyFi polymerase could amplify complicated short tandem repeat sequences (STR or SSR / microsatellite) better than any other kit.

Hiroshi Shinozuka, AgriBio, Bundoora, Australia

Product Selection

Please refer to the PCR Selection Chart to confirm the recommended product for your PCR application.

PCR can be a challenging technique, with various parameters to optimize to achieve the best results. If you are having problems, these could be easily resolved by addressing a few issues. Please see our PCR troubleshooting guide for suggestions and help with your specific problems.

These terms refer to parameters to be considered when performing PCR and are important features in selecting the correct enzyme for your needs. Understanding what they mean is therefore crucial:
Yield: The amount of DNA produced in a PCR reaction.
Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand.
Processivity: The length of time a polymerase is associated with the template and therefore the size of fragment which can be amplified.
Specificity: A measure of the unwanted by-products generated in a reaction.

During PCR setup at room temperature, standard polymerases will have some activity. When using a hot-start polymerase, this enzyme will have no activity at room temperature, thus reducing the risk of unspecific products in the reaction due to these mis-primed oligonucleotides. This feature makes this type of polymerase ideally suited to high-throughput applications, where the reactions may be left sitting at room temperature for prolonged periods of time.

In nature, the ability of a DNA polymerase to correct misincorporations of nucleotides in the DNA strand being elongated is often crucial to the survival of the host organism. This ability is termed "proofreading activity" and occurs in the 3' to 5' direction. This activity also leads to the polymerase removing unpaired nucleotides overhanging at the 3'end (A-overhangs), creating blunt ends.

All Bioline polymerases are available in the convenient format of a 2x master mix, containing all the reagents and additives necessary to perform successful PCR, with the exception of template, primers and water.
This formulation not only provides a convenient and hassle-free PCR setup, but also significantly reduces the chances of human error, inaccuracies and contamination by reducing the pipetting steps required.

If possible, we would recommend an additional cleanup of affected samples, for instance, customers had very good experiences with the clean-up of samples containing humic acids with SureClean Plus. Alternatively decreasing the template concentration will help to minimize the amount of inhibitors. The amount of primer can be increased, but do not exceed the suggested limits.

I like the Bioline MyFi polymerase, as it can amplify any sequence even if the fragment size is over 3 kb. The polymerase is also useful for cross-species amplification. The figure shows amplification results from meadow fescue using PCR primers designed for perennial ryegrass. All 5 gene sequences were successfully amplified by the cross-species PCR, even though the largest fragment (Gene 4) was over 3 kb in length.

Maiko Shi, AgriBio, Bundoora, Australia

We commonly use the Bioline MyFi polymerase, and it works better than any other polymerase kit. The MyFi polymerase could amplify complicated short tandem repeat (STR or SSR/microsatellite) sequences better than the other kits. In the figure, the MyFi kit could amplify the STR-A sequence, which the phusion kit could not. The STR-B sequence was amplified with less non-specific amplicons when the MyFi kit was used.